蚓激酶提取条件的优化与酶活性测定

蚯蚓纤溶酶是目前较为理想的溶栓药物之一,具有广泛的临床应用前景。采用盐析法从蚯蚓中提取蚓激酶,通过采用考马斯亮蓝G-250染色法间接地测定蚓激酶活性,优化出最佳的提取条件。结果显示,蚓激酶的提取最佳PH为7.5-8.0,二次盐析硫酸铵饱和度应在80%以上。     

中药"龙泉复方"的不同配伍对肿瘤细胞系HepG-2和EC-1增殖的影响

通过MTT(四甲基偶氮唑盐)法绘制细胞增殖曲线观察药物对肝癌细胞系HepG-2和食管鳞癌细胞系EC-1细胞增殖的影响,用荧光染色法进一步确定"龙泉复方"有效成分诱导细胞凋亡的较佳配伍及其有效作用浓度范围.结果表明:对抑制HepG-2细胞增殖有效作用的浓度:七味单方(马钱子、山慈菇、喜树果、龙葵、石上柏、青黛、紫菀)浸膏...     

病理组织切片中常见问题及解决方法

组织切片方法是教学、科研、病理检验工作中广泛应用的一种实验技术。文章对整个实验过程中操作程序、可能出现的问题、相关的原因及具体的解决方法进行了归纳总结,对实验步骤和操作方法等细节进行了改进,使组织固定完全,脱水彻底,透明适度,浸蜡充分,切片较薄,染色良好,为广大同行提供了一种切实可行的技术方案,并得到满意的实验结果。     

中华绒螯蟹蜕皮抑制激素多克隆抗体的制备及其表达分泌特征的组织学研究

为了探讨中华绒螯蟹(Eriocheir sinensis)蜕皮抑制激素(molt-inhibiting-hormone,MIH)的表达分泌特征,利用分子生物学技术构建了表达载体pET-30a-MIH,并制备了MIH的兔抗多克隆抗体.制备出的多克隆抗体用于MIH在中华绒螯蟹视上神经节的免疫组织化学研究.结果显示,眼柄视上神经节分泌细胞中细胞类型1,2,3,4都参与了MIH的分泌,窦腺也呈现阳性反应.     

雄性山噪鹛与白腰文鸟脑内P物质的表达差异

用免疫组织化学方法研究P物质(SP)在雄性山噪鹛与白腰文鸟听觉和发声中枢内的分布差异,结合计算机图像仪检测免疫阳性细胞和末梢的灰度值.结果:1)SP在端脑的分布为发声学习中枢X区最多,学习中枢MAN和发声高级中枢(HVc)次之,发声运动中枢古纹状体栎核最少.2)两种鸟SP的分布在听觉中枢耳蜗核、中脑背外侧核壳区、丘脑卵圆核壳区等比较相似.3)在山噪鹛发声中枢中SP的标记量比白腰文鸟丰富,尤其是在X区.结果表明:SP广泛分布于发声核团和主要听觉中枢内,提示SP可能在发声控制及听觉中枢内具有重要的生理功能,同时SP在发声中枢分布的丰富程度可能与鸣唱的复杂程度相关.     

Two forms of the membrane-bound state of the first C2 domain (C2A) of synaptotagminⅠand calcium-triggered membrane insertion

The synaptic vesicle protein synaptotagmin I(syt I) is a vesicle transmembrane protein present in synaptic vesicles, which has been proposed as the Ca^2 sensor that regulates secretion. The C2A domain is the membrane proximal part of its cytoplasmic domain. The interaction between C2A and lipid bilayer has been considered to be essential for triggering neurotransmitter release. In the present work, the measurements of membrane surface tension and surface concentration showed that the C2A domain of syt I exhibited two membrane-bound states: the surface adsorption state and the membrane insertion state. The surface absorption state formed in a Ca2~-independent manner with lower affinity, while the membrane insertion state formed with high affinity was only found in the presence of Ca^2 . Both the Ca^2 -independent and Ca^2 -dependent syt I membrane interactions required anionic phospholipids, such as phosphatidylserine (PS). When expressed into rat pheo-chromocytoma (PC12) cells and human embryonic kidney (HEK-293) cells, as demonstrated by immunofluorescence staining and subcellular fractionation, most of the C2A was found at the plasma membrane, even when the cells weredepleted of Ca^2 by incubation with EGTA. These resultssuggested a new molecular mechanism of syt I as a Ca^2 sensor in membrane fusion. Ca^2 -independent surface adsorption might attach syt I to the release site during the docking or priming step. When intracellular Ca^2 increased,syt I triggered the neurotransmitter release following the Ca^2 -dependent penetration into the target membrane.     

法夫酵母银染mRNA差异显示技术的建立

对影响银染DDRT-PCR方法的主要因素如RNA投入量、引物浓度、MgCl2浓度、dNTPs浓度以及二次扩增的退火温度进行优化.实验结果表明:适宜分析法夫酵母mRNA差异显示的银染DDRT-PCR的条件是总RNA投入量为0.4 μg、引物浓度为1.5 μmol/L、dNTPs浓度为0.25 mmol/L、MgCl2浓度为1.7 mmol/L、二次扩增的退火温度为40 ℃.     

氯化镍对绿豆和大麦核仁结构损伤的研究

应用植物A g-NOR技术,研究了不同浓度的N i2 对绿豆和大麦根细胞核仁结构的影响.结果表明:N i2 对绿豆和大麦根细胞核仁均有明显的毒害效应,用50 m g.L-1~150 m g.L-1N i2 处理绿豆和大麦根尖24 h~27 h后,细胞核仁结构发生解体,形成许多数量不等的微核仁颗粒,并从细胞核进入细胞质中,随着N i2 浓度的增加和处理时间的延长,其异常核仁的细胞比例逐渐增加.N i2 对大麦根细胞核仁的损伤效应大于绿豆.     

Mouse Karyotype Obtained by Combining DAPI Staining with Image Analysis

In this study, mitotic metaphase chromosomes in mouse were identified by a new chromosome fluorescence banding technique combining DAPI staining with image analysis. Clear 4＇, 6-diamidino-2-phenylindole （DAPI） multiple bands like （J-hands could be produced in mouse. The Meta- Morph software was then used to generate linescans of pixel intensity for the banded chromosomes from short arm to long arm. These linescans were sufficient not only to identify each individual chromosome but also analyze the physical sites of bands in chromosome. Based on the results, the clear and accurate karyotype of mouse metaphase chromosomes was established. The technique is therefore considered to he a new method for cytological studies of mouse.